JP5391282B2 - Composition for promoting hematopoietic stem cell proliferation comprising fistula extract, reishi extract and mulberry extract - Google Patents

Description

  The present invention relates to a composition for promoting the growth of hematopoietic stem cells containing a mushroom extract, and more particularly, a fistula extract, a ganoderma extract and a mulberry extract, and a plant that promotes their immune activity enhancing action. The present invention relates to a composition for promoting the growth of hematopoietic stem cells containing an extract as an effective component and a functional food.

  Immunity is large and can be divided into innate immunity from the time of birth and acquired immunity acquired by adapting to daily life. Innate immunity is also referred to as “natural immunity”, which reacts nonspecifically to antigens and has no special memory effect. Innate immune systems include skin, mucus tissue, strongly acidic gastric acid, complement in blood, etc. that block antigen invasion. Examples of the cells include macrophages responsible for phagocytosis, polymorpho nuclear leukocytes, and killer cells that can kill infected cells. In fact, many infections are protected by innate immunity. Acquired immunity is also known as “acquired immunity”. It has the feature that it memorizes the first invading antigen and reacts specifically when it re-enters, effectively removing the antigen. Fulfill. Acquired immunity is understood for convenience divided into humoral immunity and cell-mediated immunity.

  In humoral immunity, B lymphocytes recognize antigens and differentiate to secrete antibodies, which mainly have a function of removing infected bacteria. Antibodies exist in body fluids and are composed of glycoproteins called immunoglobulins (hereinafter abbreviated as Ig). There are various types such as IgG, IgM, IgM, IgA, IgD, and IgE, and each has a unique function, and some functions may overlap. IgA antibodies are characterized by being transmitted to the fetus through the placenta. Such immunity is called maternal immunity and provides protection from infection during the first few months of life. Cellular immunity plays a role in that T lymphocytes derived from the thymus recognize antigen and secrete lymphokine or kill directly infected cells. Secreted lymphokines activate macrophages and help phagocytosis. Such cellular immunity mainly has a function of removing cells infected with viruses or bacteria that grow in the cells. Acquired immunity can be obtained by vaccination with a pathogen or its toxin as an immunogen, and such immunity is called artificial immunity. E. Jenner laid the foundation of immunology by first discovering the vaccination method in this way. Various artificial immunization methods are being developed today, and their application to the prevention of cancer, acquired immune deficiency syndrome (AIDS) and hypersensitivity reactions is under study.

  Mushrooms have long been rich in flavor components and low in protein and lipids, but contain large amounts of special nutrients such as polysaccharides, vitamins and minerals, and are recognized as health foods. Pharmacological effects such as immunity enhancement and antioxidant have been clarified, and it is widely used as a material for pharmaceuticals and health functional foods.

  Conventionally, fistula fistula extract exhibits excellent activity in proliferation test and interleukin and NO (Nitric Oxide) activity in spleen and peritoneal cells, which are indicators of immunity and anticancer, and exhibits cancer cell growth inhibitory effect The effect on prevention of diseases caused by immunodeficiency and adult diseases such as cancer has been confirmed (Korea Published Patent No. 2004-0042119). In addition, it was confirmed that the composition containing ganoderma fruit body powder was excellent in immunity enhancing effect and cholesterol reducing ability, and had an effect of preventing immunity decline (Korea Published Patent No. 2003-0082619). Mulberry yellow is classified as perinus baumi in mycology, but perinus baum extract has activity against macrophages in the body, and activity of B cells in immune cells in the spleen It was confirmed that the effect of immunosuppressive activity was increased by specifically increasing the amount of the drug (Korean Patent Registration No. 0623270).

  Furthermore, it was confirmed that a complex polysaccharide cultivated by selecting and cultivating shiitake mushrooms, reishi mushrooms, fistula mushrooms, agaricus, mulberry yellow, kawaratake mushrooms and mushrooms classified as basidiomycetes enhances immune activity ( (Korean Patent Registration No. 0491362), each of which is limited to one type of mushroom, and there is a limit to making use of or improving the effectiveness of each mushroom.

  On the other hand, considering the conventional technique for enhancing the immune activity using the mushroom extract, it is only possible to enhance the immune activity through action mechanisms such as interleukin and NO activity increase, B cell proliferation in the spleen, etc. Research results that hematopoietic stem cells, which are ancestral cells of undifferentiated bone marrow hematopoietic cells that produce red blood cells, white blood cells, and platelets in the body, can be expanded to enhance immune activity are still few.

  Therefore, as a result of diligent efforts to develop a technique for activating immune effects through hematopoietic stem cell proliferation, the present inventors mixed fistula extract, reishi extract and mulberry extract, and then concentrated red ginseng. A composition prepared by adding a plant extract including a liquid, a propolis concentrate, a sagebrush extract, a goldfish extract, a licorice extract and a pig placenta concentrate to proliferate hematopoietic stem cells and promote immune cell activity As a result, the present invention has been completed.

Summary of invention

  The object of the present invention is a hematopoietic extract comprising a fistula extract, a ganoderma extract, a mulberry extract, a red ginseng concentrate, a propolis concentrate, a Chinese mugwort extract, a depression extract, a licorice extract and a pig placenta concentrate. Composition for promoting stem cell proliferation, as well as fistula extract, ganoderma extract, mulberry extract, red ginseng concentrate, propolis concentrate, Chinese mugwort extract, depression extract, licorice extract, pig placenta concentrate and It is to provide a functional food for promoting hematopoietic stem cell proliferation containing liquid fructose.

Means to achieve the task

  In order to achieve the above object, the present invention provides a fistula extract, ganoderma extract, mulberry extract, red ginseng concentrate, propolis concentrate, kawara mugwort extract, depression extract, licorice extract and pig placenta Provided is a composition for promoting hematopoietic stem cell proliferation containing a concentrate.

  The present invention also includes a fistula extract, ganoderma extract, mulberry extract, red ginseng concentrate, propolis concentrate, kawara mugwort extract, depression extract, licorice extract, pig placenta concentrate and liquid fructose. A functional food for promoting the growth of hematopoietic stem cells is provided.

  In addition, the present invention provides a fistula extract, ganoderma extract, mulberry extract, red ginseng concentrate, propolis concentrate, kawara mugi extract, urushi extract, licorice extract and pig for promoting hematopoietic stem cell proliferation. Provide use of placenta concentrate.

Furthermore, the present invention provides a fistula extract, ganoderma extract, mulberry extract, red ginseng concentrate, propolis concentrate, kawara mugwort extract, depression extract, licorice extract and pig placenta for enhancing immune activity. Provide the use of concentrate.
Other features and implementations of the invention will become more apparent from the following detailed description and the appended claims.

Detailed Description of the Invention

In one aspect, the present invention includes a fistula extract, a ganoderma extract, a mulberry extract, a red ginseng concentrate, a propolis concentrate, a sorghum extract, an umbilical extract, a licorice extract and a pig placenta concentrate. The present invention relates to a composition for promoting hematopoietic stem cell proliferation.
The present invention improves immune activity through proliferation of hematopoietic stem cells by mixing a plant extract that promotes immune cell activity enhancing action with an extract obtained by mixing a fistula extract, a ganoderma extract, and a mulberry extract. It is based on the principle that it can be enhanced.

  In the present invention, Inonotus Obliquus is a mushroom that grows infested with white birch in the northern high latitude Siberia region, and native people who drink and boil it like tea and coffee include gastrointestinal cancer patients and diabetic patients. It has been found that the active ingredients of fistula extract are polysaccharides, flavonoids, triterpenoids, inositol, chagaic acid, and alkaloids, mainly high gastrointestinal anti-ulcer activity, anti-tumor Activity, hypoglycemic and SOD activity.

  In the present invention, Ganoderma Luciderm is a mushroom that develops and grows from the base of a broad-leaved tree in the summer, which is also referred to as an immature grass, and has a diuretic, complementary liver, tonicity, mental stability, arthritis, and bronchitis. It is known to be effective in reducing blood pressure, improving hyperlipidemia, lowering blood sugar, enhancing immunity and antitumor effects.

  Mulberry yellow (Phellinus Linteus) in the present invention is a mushroom belonging to the genus Mushroom of the genus Basidiomycete tobacco, Urokotake, which is also called a wood mud mushroom. It is known that the immune function lowered after the operation is enhanced, and according to the order of this herb, it has a detoxifying action by activating uterine bleeding and substomia, irregular physiology, intestinal hemorrhage and spleen function.

  In the present invention, red ginseng is red ginseng steamed and dried from raw ginseng. Like white ginseng, glycoside, ginseng fragrance, polyacetylene-based compound, content ingredient, flavonoid, vitamin ( B group), contains trace elements, enzymes, antioxidants, organic acids and amino acids, has calming and excitatory effects on the central nervous system, and acts on the circulatory system to prevent hypertension and arteriosclerosis There is. It also lowers hematopoiesis and blood sugar levels, protects the liver, acts on the endocrine system and acts indirectly on reproductive effects, has anti-inflammatory and anti-tumor effects, has a protective effect against radiation, skin It also has the effect of protecting and softening.

  In the present invention, propolis is also referred to as Russian penicillin or natural penicillin, and has anti-inflammatory, antioxidant and immune enhancing effects, and has the most organic substances and minerals (inorganic salts), minerals, vitamins, amino acids, fats, organic acids, Flavonoids and the like play an important role in cell metabolism, and terpenes and the like have anticancer effects.

  In the present invention, the wormwood mugwort is called the four seasons mugwort or 茵 chengrass, and contains dimethyl esculetin, scopoletin, essential oil, aromatic oxycarbonic acid, flavonoid, oxycoumarin, etc. It has excellent effects in improving and treating liver function, and has an excellent effect in preventing cardiovascular diseases such as hypertension, obesity, and stroke.

  In the present invention, the gold is a dried ginger root tuber of the ginger family as it is, or steamed and dried after removing the main skin, helping the blood circulation by communicating "ki", menstrual pain, irregular menstruation, flank Remedies pain, treats vomiting, nosebleeds, hematuria, cleanses the mind, promotes bile secretion and treats gallbladder stones, contains angiogenesis inhibitors, carcinogen inhibitors Excellent function to promote bile secretion. Curcumin, a yellow pigment component of gold, is a powerful antioxidant (removing active oxygen), preventing cell oxidation and reducing inflammation, preventing dementia and delaying progression. There is also an effect.

  In the present invention, licorice is a medicinal plant.According to the East Medical Hokan, licorice is used to normalize the physiology of the eyes, nose, mouth, ears, and urine while controlling the cold and illness of the entire internal organs, and to communicate the whole blood vessel. It is known to be effective in strengthening muscles and bones, improving nutritional conditions, detoxifying the toxicity of drugs and harmonizing with each other.

  Hematopoiesis is a series of developmental processes in which a large number of mature blood cells with very specific functions and limited lifetimes are generated and continuously supplied. The hematopoietic system is composed of a very complex tissue system of hematopoietic precursor cells that can be arranged by a system of developmental capacities. The source of mature blood cells of all strains is the bone marrow, which contains cells that are in various stages of maturation, and the blood cells generated from the bone marrow are destroyed in peripheral tissues after a certain period of time. This process will occur repeatedly throughout the life of the animal. The hematopoietic action is maintained by hematopoietic stem cells (HSCs) that can proliferate and differentiate into cells of various lineages. Initially, hematopoietic stem cells are called Pluripotent Hematopoietic Stem Cells (PHSCs), which produce hematopoietic progenitor cells that can differentiate into a large variety of mature blood cells through many unexplained processes. These hematopoietic progenitor cells will then produce more differentiated cells in the next step.

  HSCs have two important characteristics. Self-renewal ability to create the same cells as self and ability to differentiate into all lineages of mature lympho-hematopoietic cells. Research on hematopoietic stem cells began when it became clear for the first time that this deficiency could be recovered by injecting normal bone marrow cells into animals suffering from bone marrow deficiency after the animals had received a sublethal dose of radiation. . From the first quantitative experiment on bone marrow recovery in such whole body irradiated mice, hematopoietic progenitor cells capable of forming bone marrow and erythroid cell colonies in the spleen and bone marrow of irradiated animals The concept was introduced. To understand the reconstitution of blood cells with self-replicating pluripotent hematopoietic stem cells (PHSCs) and the developmental biological aspects of the hematopoietic lineage, the isolation of pure pluripotent hematopoietic stem cells was required. However, since these cells exist at a very low ratio, it was difficult to separate them, but the introduction of flow cytometry and single clones for hematopoietic stem cell display antigen and differentiation antigen The development of antibodies has made it possible to select HSCs, and these properties and characteristics have also been understood.

  In the present invention, the composition for promoting hematopoietic stem cell proliferation is 5 to 50 parts by weight of Ganoderma extract, 0.5 to 30 parts by weight of mulberry extract, and red ginseng with respect to 100 parts by weight of fistula extract. 0.5-30 parts by weight of concentrate, 0.5-30 parts by weight of propolis concentrate, 0.5-30 parts by weight of wormwood extract, 0.5-50 parts by weight of extract of licorice, 0.5-0.5 parts of licorice extract You may contain 50 weight part and 0.5-30 weight part of pig placenta concentrate.

  In the present invention, when fistula extract (2% solid content) is taken 3 times a day, 8 mL at a time, gastric polyposis patients are remarkably improved within one month, and gastric ulcer patients are also painful within one month. Vomiting and belching disappeared. Since fistula shows SOD activity (ability to remove active oxygen) more than 3 times higher than general mushrooms, taking a fistula eliminates the active oxygen that causes in vivo damage, and all cells in the body Various bioactive substances were isolated and purified from mushrooms, and among them, beta-glucan, glycoprotein, dietary fiber, antioxidants, etc. were confirmed to be the main components representing their bioactivity. However, the content of beta glucan (57 μg / 10 g), which has an immune enhancing effect, in fistula fistula is more than 10 times higher than the content of beta glucan in mulberry yellow (5 μg / 10 g), and the SOD activity is also a general mushroom. 3 times higher than Accordingly, the fistula occupies the highest content in the present composition in order to promote the proliferation or activity of hematopoietic stem cells that are ancestral cells of immune cells.

  In the present invention, the ganoderma extract is 5 to 50 parts by weight, preferably 5 to 15 parts by weight, with respect to 100 parts by weight of the fistula moth extract, If it is out of the range, the amount of hematopoietic stem cell proliferation is very small.

  In the present invention, it has been confirmed that Ganoderma extract protein polysaccharide significantly increases the ability of B cells to produce antibodies and enhances the immune response of T cells in connection with the differentiation and proliferation of hematopoietic stem cells. It has been confirmed that protein polysaccharides extracted from Ganoderma mycelium have an inhibitory effect on pulmonary metastasis of mice by intraperitoneal administration, and this effect is due to the promotion of interleukin secretion by activation of macrophages and NK cells. Was analyzed.

  In the present invention, the mulberry yellow extract is 0.5 to 30 parts by weight, preferably 0.5 to 10 parts by weight, based on 100 parts by weight of the fistula extract. In the case where the content of P is out of the above range, the activity of T cells becomes weak, and the activity and immune activity of hematopoietic stem cells decrease.

  In the present invention, it has been reported that mulberry yellow extract improves immunity through the activity of NK cells and activates bone marrow-derived dendritic cells to increase the number of helper T cells and to exhibit an anticancer effect. Mulberry yellow hot water extract was confirmed to show an anticancer effect by significantly increasing the activity of B cells producing antibodies and also increasing the activity of macrophages to induce NO production.

  In the present invention, the red ginseng concentrate is 0.5 to 30 parts by weight, preferably 1 to 15 parts by weight, based on 100 parts by weight of the fistula extract, When the content of is out of the above range, the hematopoietic stem cell activity falls, and the immune activity is weakened by the decrease in T cell activity.

  In the present invention, the propolis concentrate is 0.5 to 30 parts by weight, preferably 0.5 to 10 parts by weight, based on 100 parts by weight of the fistula extract, and the propolis concentrate Antioxidant and antibacterial effects are reduced when the content of is out of the above range.

  In the present invention, the wormwood extract is 0.5 to 30 parts by weight, preferably 1 to 20 parts by weight, based on 100 parts by weight of the fistula extract, and contains the wormwood extract When the amount is out of the above range, the cell activity decreases and the detoxification action decreases.

  In the present invention, the gold extract is 0.5 to 50 parts by weight, preferably 1 to 30 parts by weight with respect to 100 parts by weight of the fistula extract, and contains the gold extract. When the amount is out of the above range, the activity of hematopoietic stem cells decreases due to a decrease in antioxidant activity.

  In the present invention, the licorice extract is 0.5 to 50 parts by weight, preferably 0.5 to 20 parts by weight, based on 100 parts by weight of the fistula extract, In the case where the content of P is out of the above range, the anti-inflammatory action and liver detoxification action are weakened.

  In the present invention, the pig placenta concentrate is 0.5 to 30 parts by weight, preferably 1 to 20 parts by weight, based on 100 parts by weight of the fistula extract, When the content of is out of the above range, the cell growth promoting activity falls.

  In the present invention, red ginseng is effective in enhancing immunity, recovering from fatigue, and improving blood flow, propolis is antibacterial and antioxidative, Kawara mugwort promotes cell growth and improves liver function, and gold is anti It is effective in suppressing the occurrence of oxidation and cancer. Licorice has liver detoxification and antibacterial effects, and pig placenta promotes cell growth, and has immunity enhancement and anti-aging effects. Accordingly, these components are added to the mushroom extract and provided as a composition for promoting hematopoietic stem cell proliferation, which has excellent effects of enhancing immunity and proliferating hematopoietic stem cells.

  In the present invention, fistula moth is extracted with purified water at 50 to 60 ° C. for 4 to 8 hours, Ganoderma and mulberry are extracted with purified water at 70 to 80 ° C. for 8 to 16 hours and filtered through a 1 to 10 μm filter. Then, the step of concentrating under reduced pressure at 60 to 80 ° C. and mixing purified water with Kojiro extract, Ganoderma extract and Mulberry extract, Red ginseng concentrate, propolis concentrate, and porcine placenta concentrate by adding concentrated solution that was extracted with purified water at 70-80 ° C for 8-16 hours, filtered through a 1-10 µm filter, and decompressed at 60-80 ° C. Can be added to produce a composition for promoting hematopoietic stem cell proliferation.

  The composition for promoting hematopoietic stem cell proliferation of the present invention can be provided in the form of a pharmaceutical composition or a food composition.

  Pharmaceutical composition comprising fistula extract, ganoderma extract, mulberry extract, red ginseng concentrate, propolis concentrate, cormorant mugwort extract, garnet extract, licorice extract and pig placenta concentrate according to the present invention May contain additional suitable carriers or excipients according to conventional methods, additionally mixed or diluted with a diluent. Suitable carriers, excipients and diluents include, for example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil. The composition may additionally contain fillers, anti-aggregating agents, lubricants, wetting agents, perfumes, emulsifiers, preservatives and the like. The composition of the present invention is formulated in a manner well known in the art so that it can provide rapid or delayed release of the active ingredient after administration to a mammal. The dosage forms may be in the form of purified, powder, pills, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions and sterile powders and the like. The compositions of the invention can be administered via a variety of routes including oral, transdermal, subcutaneous, intravenous, or muscle. The dose of the composition containing the extract according to the present invention to the human body varies depending on the age, weight, sex, dosage form, health condition, and degree of disease of the patient. It is ˜240 mL / day, desirably 80 to 160 mL / day, and may be administered once to three times a day at regular time intervals according to the judgment of a doctor or pharmacist. It can be increased or decreased according to age, sex, weight, and health status.

  The food composition of the present invention can be produced in various food forms, for example, in the form of a beverage composition, and may be produced containing liquid fructose.

  Furthermore, the present invention provides, in another aspect, a fistula extract, a ganoderma extract, a mulberry extract, a red ginseng concentrate, a propolis concentrate, a Chinese mugwort extract, a depression extract, a licorice extract, and a pig placenta concentrate. The present invention relates to a functional food for promoting hematopoietic stem cell proliferation containing liquid and liquid fructose.

  In the present invention, “functional food” is the same term as food for special health use (FoSHU), and is processed to efficiently exhibit a bioregulatory function in addition to nutrition supply. Means medicine, food with high medical effect. In the present invention, “health food” means a food having an active health maintenance and promotion effect as compared with general food, and “health supplement food” means health supplement. Means food intended for. In some cases, the terms functional food, health food, health supplement are used interchangeably. The food is produced in various forms in order to obtain a useful effect for promoting hematopoietic stem cell proliferation.

  The present invention relates to 5 to 50 parts by weight of Ganoderma extract, 0.5 to 30 parts by weight of mulberry yellow extract, 0.5 to 30 parts by weight of red ginseng concentrate, Propolis concentrate 0.5-30 parts by weight, Kawara mugwort extract 0.5-30 parts by weight, metallurgical extract 0.5-50 parts by weight, licorice extract 0.5-50 parts by weight, pig placenta concentrate 0. The present invention relates to a functional food for promoting hematopoietic stem cell proliferation containing 5 to 30 parts by weight and liquid fructose 20 to 80 parts by weight.

  In the present invention, the liquid fructose is 20 to 80 parts by weight, preferably 30 to 60 parts by weight with respect to 100 parts by weight of the fistula extract, and the liquid fructose content is in the above range. If it falls outside, the preference for the taste as a beverage product is reduced.

Hereinafter, the present invention will be described in more detail through examples. It will be apparent to those skilled in the art that this example is merely illustrative of the invention and that the scope of the invention is not to be construed as limited by this example.
[Example 1] Manufacture of a composition beverage for promoting hematopoietic stem cell proliferation A fistula (Russia) was extracted with purified water (weight 10 times the dry weight of mushrooms) at 50 to 60 ° C for 4 to 8 hours. (Yeoju, Korea) and Mulberry (Towon Agricultural, Korea) were extracted with purified water at 70-80 ° C (10 times the dry weight of mushrooms) for 8-16 hours and filtered through 1-10 µm filter. The solution was concentrated under reduced pressure at 60 to 80 ° C., and purified water was extracted with 15 g of fistula extract (solid content 5%), ganoderma extract (solid content 5%) 1.3 g, mulberry yellow extract (solid content 5%) ) Mixed to 0.7 g, and extracted Kawara mugwort, goldfish and licorice with purified water at 70-80 ° C. for 8-16 hours, filtered through 1-10 μm filter, and concentrated under reduced pressure at 60-80 ° C. 0.6 g of extract (5% solids), 1.0 g of gold extract (5% solids), sweet Add 1.0 g of extract (solid content 32%) and supplementarily red ginseng concentrate (saponin 70 mg / g) 0.6 g, propolis concentrate (flavonoid 1%) 0.3 g, pig placenta concentrate (solid content 55%) 0.6 g and liquid fructose (F-55) 7.5 g were added to produce a hematopoietic stem cell proliferation promoting composition beverage (Table 1).

[Comparative Example 1] Placebo Beverage Production A placebo beverage was produced by adding 30 g of Kawaramugi extract (solid content 1.5%) and 12.5 g of liquid fructose (F-55) to 57.5 g of purified water (Table 2). ).

[Experimental Example 1] Observation of immunostimulatory effect of composition for promoting hematopoietic stem cell proliferation The composition beverage for promoting hematopoietic stem cell proliferation produced in Example 1 and the placebo beverage produced in Comparative Example 1 were used for subjects. Then, hematopoietic stem cell proliferation effect experiment was carried out.

  The test subjects were healthy volunteers, general men and women in their 40s and 70s, and blood collected from subjects using TBA-20R (Toshiba, Japan), an automatic serum analyzer, one week before entering the test. GOT and GPT values were measured, blood analysis was performed, and severe liver disease subjects with high GPT and GOT values were excluded from the experiment. After the blood analysis, the composition for promoting hematopoietic stem cell proliferation and the placebo drink taking group were randomly classified into 27 and 12 respectively for all 39 selected persons, and then the composition for promoting hematopoietic stem cell proliferation Physical drinks and placebo drinks were provided separately and taken for 4 weeks. The sex composition is 33 females and 6 males, each subject takes 80 mL of a composition for promoting hematopoietic stem cell proliferation once a day in the morning and evening twice a day, and a placebo drink in the same manner. Ingested for a total of 4 weeks. The subjects were instructed to take only the composition drink for promoting hematopoietic stem cell proliferation and the placebo drink used for the test for 4 weeks, and other health foods and alcohol were strictly restricted. Table 3 shows the physical characteristics of the hematopoietic stem cell proliferation promoting composition drink group and the placebo drink group.

  In the following, the materials of this study calculated the mean and standard deviation for each variable, and the material analysis used the SPSS 16.0 statistical program. In order to compare the difference between the placebo drink and the composition drink for promoting hematopoietic stem cell proliferation against the measured cell variable (before taking, 2 weeks after taking, 4 weeks after taking), it was analyzed by applying pared t-test. . And in order to compare the difference between the groups by the placebo drink and the composition drink use for hematopoietic stem cell proliferation promotion, it analyzed by applying independent t-test. The statistical significance level was p <0.05.

1-1: Change in the number of hematopoietic stem cells Peripheral blood leukocyte samples were prepared in a single cell suspension of 1 × 10 ⁶ cells / mL. Place 100 μL of white blood cell sample and 10 μL of CD45-FITC in the control tube, and add 100 μL of white blood cell sample, 10 μL of CD45-FITC and 10 μL of CD34-PE to the test tube, then mix and react for 15 minutes in the dark room at room temperature. did. Each tube was charged with 2 mL of Lysing Solution (1 ×, BD, USA) and immediately mixed well, and then reacted in a dark room at room temperature for 10 minutes. Each reaction sample was centrifuged at room temperature for 5 minutes (X 300 g) and the upper layer solution was removed. Then, 2 mL of PBS was added and mixed well, followed by centrifugation for 5 minutes (X 200 g). The upper layer solution of the tube was removed, and 0.5 mL of PBS (with 0.5% paraformaldehyde) was added and stained.

Using flow cytometry FACSCalibur and Cell Quest (trade name) program (Becton Dickenson, USA), cells that do not express CD45 in SSC (side scatter) vs CD45 FL-1 mode by ISHAGE method or dead In addition, after selecting all cells expressing CD45 antigen, cells expressing CD34 antigen were selected using SSC vs CD34 FL-2 mode. Among these cells, SSC vs CD45 F-L1 and FSC (forward scatter) vs SSC mode have low SSC and low CD45 FL-1 and FSC in order to prevent contamination of cells showing non-specific reactions. Only moderate or moderate cells were selected. 100,000 CD45 ⁺ cells were analyzed, and the CD34 ⁺ cell ratio (%) was determined by the formula “(CD34 ⁺ , low SSC, low to intermediate FSC and CD45 FL-1 cells / CD45 ⁺ cells) × 100” The number of CD34 ⁺ cells was calculated by the formula “white blood cell count × CD34 (%)”.

  In order to separate peripheral blood leukocytes, 10 mL of D-PBS was diluted with 10 mL of peripheral blood collected from the subject's antebrachial vein. Put 15 mL of Ficoll-Plaque (trade name) plus (GE Healthcare) in a 50 mL Conical tube, slowly add 20 mL of diluted blood on it, then centrifuge for 50 minutes (X700 g) and only the buffy coat formed in the upper layer Was collected and washed twice with D-PBS to separate peripheral blood leukocytes. The isolated leukocytes were observed for changes in the number of hematopoietic stem cells before and after taking the hematopoietic stem cell proliferation promoting composition drink group and the placebo drink group.

  As a result, hematopoietic stem cell proliferation-promoting composition drink group, the average number of hematopoietic stem cells was 3.89 per 100,000 total leukocytes in peripheral blood before taking from 8.78 after 2 weeks and 4 weeks after taking ( p <0.01) and 7.14 (p <0.001) markedly increased, showing a statistically significant difference. However, in the placebo drink group, hematopoietic stem cells average 2.67 per 100,000 total leukocytes in peripheral blood before taking from 2.75 after taking 2 weeks and 4 weeks after taking (p <0.01). It decreased significantly to 4.38 (p <0.05). On the other hand, as a result of applying the independent t-test and analyzing the hematopoietic stem cell change for each dosing period between the placebo drink group and the hematopoietic stem cell proliferation promoting composition drink group, it was already significant between the two groups before taking A difference (p <0.05) is shown, and it is difficult to recognize the dominant difference between the two groups shown 2 weeks and 4 weeks after the dose. Hematopoietic stem cell proliferation-promoting composition drink group, 4 weeks after taking, 15 out of 27 people increased the number of hematopoietic stem cells, 7 decreased, 5 people showed little change there were. In addition, one out of 27 women in the group taking hematopoietic stem cell proliferation-promoting composition beverage experienced mild abdominal pain at the beginning of drinking, and no other side effects appeared.

1-2: Change in the number of lymphocytes (Lymphocyte) Beckman Coulter STKS (Beckman, USA), an automatic blood cell analyzer, was used to measure the number of lymphocytes in blood collected from subjects. In the group of hematopoietic stem cell proliferation promoting composition drinks, the average decreased from 1,850 cells / μL before administration to 1,780 cells / μL and 1,720 cells / μL after 2 weeks and 4 weeks, but statistically There was no significant difference. There was no difference in the number of lymphocyte cells in the placebo drink group after 2 weeks and 4 weeks (Table 5).

1-3: Change in T cell subtype (CD4 ⁺ , CD8 ⁺ , CD4 ⁺ / CD8 ⁺ ) ratio Peripheral blood leukocyte samples were prepared in a single cell suspension at 1 × 10 ⁶ cells / mL, and leukocytes were prepared. After adding paraformaldehyde to the sample at 1% concentration and fixing at 4 ° C. for 1 hour, normal goat serum (Vector, USA) was 2.5% and normal horse serum (Vector, USA) at 2.5% concentration, respectively. The mixture was added and reacted at 4 ° C. for 1 hour. The reacted sample was separated into a 96-well plate by 40 μL per well. 4 μL of CD3 antibody (BD, USA) was added to each well in which the sample was separated. 4 μL each of CD4 (BD, USA) antibody and CD8 (BD, USA) antibody was placed in wells labeled with CD4 and CD8, and double-stained at 4 ° C. for 1 hour. Furthermore, it was washed 3 times with DPBS. Each reaction sample was transferred to a FACS tube and stained.

Using flow cytometry FACSCalibur and Cell Quest (trade name) program (Becton Dickenson, USA), CD3 FL-2 (x axis) versus CD4 FL-1 (y axis), CD8 FL-4 (y The axis was divided into quadrants based on the negative control density plot. Analysis for each CD3 / CD4 and CD3 / CD8 was performed, and the ratio of helper T cells that were CD3 ⁺ / CD4 ⁺ took the values shown on the 2/4 plane. In the same way, the ratio of cytotoxic T cells that are CD3 ⁺ / CD8 ⁺ also took the values shown on the 2/4 plane. The ratio of CD4: CD8 was analyzed using the ratio of CD4 ⁺ cells and CD8 ⁺ cells analyzed in this way.

1) CD4 ⁺ T cells (helper T cells)
CD4 ⁺ T cells showed no difference in the group taking the composition for promoting hematopoietic stem cell proliferation from 42.35% before taking to 42.41% after taking 4 weeks. There was no significant difference in changes in the placebo drink group even before and after the dose. There was no difference in the change in Cd4 ⁺ T cells for each dosing period between the placebo drink group and the hematopoietic stem cell proliferation promoting composition drink group (Table 6).

2) CD8 ⁺ T cells (cytotoxic T cells)
CD8 ⁺ T cells markedly increased from 22.25% before taking to 24.08% after taking 4 weeks after taking the composition for promoting hematopoietic stem cell proliferation (p <0.05), and even in the placebo drink taking group. It markedly increased from 19.88% before taking to 23.14% after 4 weeks of taking (p <0.01). It is difficult to recognize that CD8 ⁺ T cells are statistically different due to the effect of taking a composition beverage for promoting hematopoietic stem cell proliferation (Table 6). There was no difference in the number of CD8 ⁺ T cells in each dose period between the placebo drink group and the hematopoietic stem cell proliferation promoting composition drink group (Table 6).

3) Change in CD4 ⁺ / CD8 ⁺ ratio (%) The CD4 ⁺ / CD8 ⁺ ratio was 2 in the group of hematopoietic stem cell proliferation-promoting composition drinks taken from 2.21% before taking 2 weeks and 4 weeks after taking each. .16, 2.06, but there was no statistically significant difference. In the placebo drink group, it decreased from 2.53 before taking to 2.32 and 2.19 after 2 weeks and 4 weeks respectively, but there was no difference. The CD4 ⁺ / CD8 ⁺ T cell ratio is utilized as an important index indicating cellular immune function, and the normal function is about 1.8 to 2.2 and decreases to 1.5 or less. Falling and viral infection may increase. It was revealed that in the group of hematopoietic stem cell proliferation promoting composition drinks in this study, the CD4 ⁺ / CD8 ⁺ T cell ratio was 2.0 or more before and after taking, and the normal values were maintained (Table 6). . When such a result is seen, taking the composition drink for hematopoietic stem cell proliferation promotion for 4 weeks seems to have a positive influence on the immune function of T cells.

1-4: Change in NK cell ratio (%) A peripheral blood leukocyte sample was prepared in a single cell suspension of 1 × 10 ⁶ cells / mL, paraformaldehyde was added to the leukocyte sample at a concentration of 1%, After fixing at 4 ° C. for 1 hour, normal goat serum (Vector, USA) was added at 2.5% and normal horse serum (Vector, USA) at 2.5% concentrations, respectively, and reacted at 4 ° C. for 1 hour. The reacted sample was stocked in a 96-well plate at 40 μL per well. 4 μL of CD3 antibody (BD, USA) was added to each well in which the sample was separated. CD56 antibody (BD, USA) was placed in wells labeled with CD56 in 4 μL, and double-stained at 4 ° C. for 1 hour. Furthermore, it was washed 3 times with DPBS. Each reaction sample was transferred to a FACS tube and stained.

Using the flow cytometry FACSCalibur and Cell Quest program (Becton Dickenson, USA) to analyze the ratio of NK cells, CD56 FL-4 (y axis) to CD3 FL-2 (x axis) The axis was divided into quadrants based on the negative control density plot. Samples that were immunofluorescently stained with CD3 / CD56 were analyzed, and the ratio of natural killer cells that were CD3 ⁻ / CD56 ⁺ took the values shown on the quarter plane.

The ratio of NK cells (CD3 ⁻ , CD56 ⁺ ) in lymphocytes of peripheral blood decreased from 17.54% before taking the composition for promoting hematopoietic stem cell proliferation to 17.01% after taking 4 weeks. There was no significant difference. It was found that NK cells decreased after 4 weeks of taking in the placebo drink group compared with before taking the placebo drink, but there was no significant difference (Table 7).

[Comparative Example 2] Comparison of immune enhancement mechanism Mechanism of immune activity enhancement of a composition beverage for promoting hematopoietic stem cell proliferation containing the fistula extract, reishi extract and mulberry extract of Example 1 in the prior art As a result, Korean Patent No. 2004-0042119 is superior in proliferation test, interleukin, NO activity and cancer cell proliferation inhibitory effect in spleen and peritoneal cells using fistula extract, Published Patent No. 2003-0082619 is excellent in enhancing immunity and cholesterol-reducing ability by ganoderma and is excellent in preventing immunity reduction. In Korean Patent Registration No. 623270, mulberry yellow extract is present in immune cells in the spleen. Although it has been reported that the activity of the B cell is specifically increased to have an effect on the immune activity, there has not yet been reported on the enhancement of the immune activity by the proliferation of hematopoietic stem cells.

  On the other hand, as a result of considering before and after taking the composition for promoting hematopoietic stem cell proliferation containing the fistula extract, ganoderma extract and mulberry extract extracted in Example 1 according to the present invention, Significantly increased the value of hematopoietic stem cells, which helped to enhance overall immune activity by normalizing whole blood cells, and was confirmed to be superior to conventional single mushroom extracts ( Table 8).

  The composition and functional food according to the present invention remarkably enhance the proliferation effect of hematopoietic stem cells, assist immune activity, and anti-cancer, anti-inflammatory, cell activity promotion, damaged tissue accompanying hematopoietic stem cell proliferation and immune activity enhancement Positive effect on regeneration and prevention of cardiovascular disease.

  Although specific portions of the subject matter of the present invention have been described in detail above, such specific notation is merely a preferred embodiment for those having ordinary skill in the art, thereby Obviously, the scope of the present invention is not limited. Accordingly, the substantial scope of the present invention is defined by the appended claims and equivalents thereof.

Claims (2)

  A composition for promoting hematopoietic stem cell proliferation, comprising a fistula extract, a ganoderma extract, a mulberry extract, a red ginseng concentrate, a propolis concentrate, a cormorant mugwort extract, a goldfish extract, a licorice extract and a pig placenta concentrate object.   5 to 50 parts by weight of Ganoderma extract, 0.5 to 30 parts by weight of mulberry extract, 0.5 to 30 parts by weight of red ginseng concentrate, 0 to propolis concentrate .5-30 parts by weight, Kawara mugwort extract 0.5-30 parts by weight, goldfish extract 0.5-50 parts by weight, licorice extract 0.5-50 parts by weight and pig placenta concentrate 0.5-30 parts by weight The composition for promoting hematopoietic stem cell proliferation according to claim 1, comprising a part.